Decellularized Matrix Preparation for Low-Immunogenic ECM Scaffolds
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Solution Overview
Problem
Existing methods for preparing decellularized matrix biomaterials face challenges in effectively removing immunogenic ingredients while minimizing damage to the extracellular matrix (ECM) structure and retaining bioactive ingredients, leading to potential immune reactions and reduced tissue regeneration capabilities.
Innovation Solution
A method involving pretreating, degreasing, disinfecting, decellularizing, removing α-Gal antigen, and sterilizing animal tissues using specific reagents and enzymes to achieve efficient removal of immunogenic ingredients while preserving the ECM's structural integrity and bioactive components like collagen, glycosaminoglycan, and laminin.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If strong detergents (SDS, Triton X-100, CHAPS) are used for decellularizing, then decellularizing effect is improved, but three-dimensional structure of ECM is damaged and bioactive ingredients are lost
Solution Approach 1:
The patent uses a mild non-ionic detergent (Tween-20) as an intermediary decellularizing agent that effectively removes cells while preserving the ECM structure. This mediator achieves decellularization without the harsh effects of strong detergents, maintaining the integrity of collagen and other bioactive components.
Solution Approach 2:
The patent changes the parameters of the decellularizing solution by using a mild non-ionic detergent with specific concentration ranges (0.1-1% Tween-20) and controlling treatment time (6-48 hours). This parameter optimization allows effective decellularization while minimizing damage to the ECM three-dimensional structure.
2Reliability
If NaOH is used for decellularizing, then decellularizing effect is improved, but collagen structure of ECM is damaged and growth factors are removed
Solution Approach 1:
The patent replaces the harsh NaOH intermediary with a mild non-ionic detergent (Tween-20) that acts as a gentler mediator for cell removal. This intermediary achieves decellularization while preserving collagen structure and growth factors, avoiding the damaging effects of strong alkaline reagents.
Solution Approach 2:
The patent changes the chemical parameters of the decellularizing agent from strong alkaline (NaOH) to mild non-ionic detergent (Tween-20) with controlled concentration and treatment time. This parameter change maintains decellularizing effectiveness while preserving ECM structural integrity.
3Reliability
If trypsin is used for decellularizing, then decellularizing effect is improved, but ultrastructure of ECM is damaged and mechanical properties decrease
Solution Approach 1:
The patent replaces the proteolytic enzyme trypsin with a non-ionic detergent (Tween-20) as an intermediary decellularizing agent. This mediator removes cells through solubilization rather than proteolysis, preserving the ultrastructure and mechanical properties of the ECM.
Solution Approach 2:
The patent substitutes the biochemical mechanism of trypsin (proteolytic digestion) with a physical-chemical mechanism (non-ionic detergent solubilization). This substitution removes cells while preserving the structural integrity and mechanical properties of collagen and other ECM components.
4Reliability
If neutral protease is used for decellularizing, then decellularizing effect is improved, but ultrastructure of ECM is damaged and fibronectin and collagen IV are degraded
Solution Approach 1:
The patent replaces neutral protease with a non-ionic detergent (Tween-20) as an intermediary decellularizing agent. This mediator removes cells without proteolytic activity, thereby preserving the ultrastructure and specific ECM proteins such as fibronectin and collagen IV.
Solution Approach 2:
The patent substitutes the enzymatic mechanism of neutral protease with a non-enzymatic detergent-based mechanism. This substitution achieves decellularization while preserving the ultrastructure and specific ECM proteins that would otherwise be degraded by proteolytic enzymes.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method results in a decellularized matrix biomaterial with low immunogenicity and high bioactivity, supporting cell adhesion, proliferation, and tissue regeneration, with improved biocompatibility and structural stability.
Implementation Method 1
decellularizing: mixing the matrix material with a solution of 0.01-0.5 wt % alkaline protease at a mass ratio of matrix material to solution of 1:3-10 and shaking for 60-240 min
Implementation Method 2
degreasing: mixing the matrix material with a degreasing reagent according to a mass ratio of matrix material to solution of 1:5-20 and shaking for 30-180 min; wherein the degreasing reagent is at least one selected from the group consisting of a solution of 2-10 wt % sodium dodecyl sulfonate
Implementation Method 3
disinfecting: mixing the matrix material with a disinfectant according to a mass ratio of matrix material to solution of 1:3-10 and shaking for 30-180 min; wherein the disinfectant is selected from the group consisting of a mixed solution of 0.02-0.2 wt % peroxyacetic acid and 10-40 wt % ethanol
Data Source
AI summary
A preparation method for a decellularized matrix biomaterial, which combines specific pretreatment, degreasing, disinfection, decellularization, α-Gal antigen removal and sterilization, and reserves the integrity of bioactive components and structures in a material to the maximum extent while effectively removing an antigenic substance of an animal tissue, so that a decellularized matrix biomaterial having low immunogenicity and high biological activity is obtained. In addition, the decellularized matrix biomaterial prepared by the method effectively reduces immunogen components, has good biocompatibility and biological safety, is beneficial to cell adhesion, aggregation and proliferation on the surface, and can be used as a high-activity biological matrix material to be applied to skin soft tissue defect, gingival recession and periodontal soft tissue repair.